Endothelin A receptor blockade improves regression of flow-induced pulmonary vasculopathy in piglets  Olaf Mercier, MD, Edouard Sage, MD, Mohammed Izziki,

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Endothelin A receptor blockade improves regression of flow-induced pulmonary vasculopathy in piglets  Olaf Mercier, MD, Edouard Sage, MD, Mohammed Izziki, PhD, Marc Humbert, MD, PhD, Philippe Dartevelle, MD, Saadia Eddahibi, PhD, Elie Fadel, MD, PhD  The Journal of Thoracic and Cardiovascular Surgery  Volume 140, Issue 3, Pages 677-683 (September 2010) DOI: 10.1016/j.jtcvs.2010.01.004 Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Box plot of the percentage of media thickness of small pulmonary arteries (<200 μm) assessed by morphometry (×400) in the sham group (35.5% ± 0.9%), high-flow group (54.9% ± 1.3%), flow-correction group (50.3% ± 1.3%), and flow-correction plus anti-ETA group (33.2% ± 8%). Media thickness returned to sham values 1 week after shunt closure in the group given anti-ETA therapy. MT, Media thickness; ED, external diameter; ID, internal diameter. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 677-683DOI: (10.1016/j.jtcvs.2010.01.004) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Evaluation of in situ proliferation of pulmonary artery smooth muscle cells (PA-SMCs) by PCNA labeling (A, B, C, D) of PA-SMC death by TUNEL (E, F, G, H) and of collagen accumulation by Masson trichrome staining (I, J, K, L) in the high-flow, flow-correction, flow-correction plus anti-ETA, and sham groups (×400). Positive cells by TUNEL and PCNA labeling are brown. Collagen is stained blue by Masson's trichrome. Pulmonary artery medial hypertrophy in the high-flow group is related to PA-SMC proliferation (B, arrow) without intravascular collagen accumulation (J). The decrease in media thickness 1 week after shunt closure alone or with anti-ETA was associated with a decrease in PCNA-positive cells (C, D) and with an increase in TUNEL-positive cells (G, H, asterisk). Anti-ETA therapy was started 1 week before shunt closure and the smaller media thickness value in the group given anti-ETA indicates that the treatment was effective in diminishing the medial hypertrophy. TUNEL, Terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling; ETA, endothelin receptor type A; PCNA, proliferating cell nuclear antigen. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 677-683DOI: (10.1016/j.jtcvs.2010.01.004) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Percentage of PCNA and TUNEL-positive cells in the distal pulmonary artery wall in all groups of piglets. PA-SMC proliferation was noted in the high-flow group but was no longer present 1 week after shunt closure. PA-SMC apoptosis was increased 1 week after shunt closure with or without anti-ETA therapy. TUNEL, Terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling; ETA, endothelin receptor type A; PCNA, proliferating cell nuclear antigen. P < .05 vs sham. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 677-683DOI: (10.1016/j.jtcvs.2010.01.004) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Quantification by RT-PCR of the mRNAs for endothelin-1 (ET-1) and its receptors (ETA and ETB) in lung parenchyma. High blood flow in the lungs increased the expression of ET-1 and ETA mRNAs but did not change the steady-state level of ETB mRNA expression. One week after shunt closure, ET-1 and ETA mRNA levels had returned to normal. Except for ET-1, mRNA levels decreased similarly after flow correction with and without anti-ETA therapy. The ET-1 level remained elevated in the anti-ETA group. ∗P < .05 vs sham. ∗∗P< .05 vs 1 week shunt-closed. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 677-683DOI: (10.1016/j.jtcvs.2010.01.004) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Quantification of angiopoietin-1 (Ang-1) and Tie 2 mRNA levels in lung parenchyma from the sham, high-flow, flow-correction, and flow-correction plus anti-ETA groups. High blood flow in the lungs increased the expression of angiopoietin-1 and Tie2 mRNAs. Shunt closure with or without anti-ETA reversed the overexpression of these genes. ETA, Endothelin receptor type A. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 677-683DOI: (10.1016/j.jtcvs.2010.01.004) Copyright © 2010 The American Association for Thoracic Surgery Terms and Conditions